12,17-dioxy and 12-oxo-17-oxy-a-norprogesterones

ABSTRACT

12-17-dioxy and 12-oxo-17-oxy-A-norprogesterones which are useful as anti-androgenic agents.

United Siaies Pmem Levine et all.

12,17-DIOXY AND l2-OX0-l7-0XY-A- NORPROGESTERONES Inventors: Seymour D. Levine, Princeton; Pacifio A. Principe, South River,

both of NJ.

Assignee: E. R. Squibb & Sons, Inc, New

York, N.Y.

Filed: June 25, 1965 Appl. No.1 467,103

US. Cl. .260/488 B, 195/51, 260/410, 260/468 R, 260/476 C, 260/486 R, 260/586 H, 260/999 lint. Cl. ....C07c 49/44, C070 69/ 14, C07c 69/16 [451 Mar. 27, 1973 [58] Field of Search ..260/488 B, 586 H, 410, 486,

Primary Examiner-Vivian Garner Attorney-Lawrence S. Levinson and Merle J. Smith ABSTRACT 1 2-1 7-dioxy and 12-oxol 7-oxy-A-norprogesterones which are useful as anti-androgenic agents.

5 Claims, No Drawings wherein R and R are selected from the group consisting of hydrogen and acyl.

The preferred acyl radicals are those of hydrocarbon carboxylic acids of less than twelve carbon atoms, as exemplified by the lower alkanoic acids (e.g., acetic, propionic butyric and tert-pentanoic acid), the lower alkenoic acids, the monocyclic aryl carboxylic acids (e.g., benzoic and toluic acids), the monocyclic aryl lower alkanoic acids (e.g., phenacetic and fl-phenylpropionic acid), the cycloalkane carboxylic acids and the cycloalkene carboxylic acids.

The novel compounds of this invention are pharmacologically active substances which possess anti-androgenic activity (i.'e., they inhibit the actions of androgens),'and which may be used in the treatment of such conditions as hyperandrogenic acne.

The compounds may be formulated for such administration, the concentration and/or dosage being based on the activity of the particular compound and the requirements of the patient.

The final products of the instant invention may be prepared by the processes of this invention which entails a number of steps beginning with l7a-hydroxy-A- norprogesterone as starting material. The preparation of this compound is disclosed in copending U. S. Pat. application, Ser. No. 399,838, filed Sept. 28, 1964, now U.S. Pat. No. 3,381,029. The process of the instant invention may be represented by the following equations wherein R and RI areashereinbefore defined:

CH; CH:

the starting material by subjecting the latter to the action of a microorganism of the genus Corticium or to the action of the enzymes thereof under oxidizing and preferably aerobic conditions.

To prepare the compounds of this invention, the starting reactant may be first subjected to the action of enzymes of a microorganism of the genus Corticium under oxidizing conditions. This oxidation can best be effected either by including the starting reactant in an aerobic culture of the microorganism, or by bringing together in an aqueous medium, the compounds, air, and enzymes of non-proliferating cells of the microorganism.

In general, the conditions of culturingthe microorganism for the purposes of this invention are (except for the inclusion of the starting reactant to be converted), the same as those of culturing various other microorganisms for the production of antibiotics, and other like substances. The microorganism is grown aerobically in contact with, (in or on) suitable fermentation medium. A suitable medium essentially comprises a source of carbon and energy. The latter may be a carbohydrate, for example, molasses, glucose, maltose, starch or dextrin, a fatty acid, a fat and/or the compound itself. Preferably, however, the medium includes an assimilable source of carbon and energy in addition to the steroid. Among the fats utilizable for the purpose of this invention are lard oil, soybean oil, linseed oil, cottonseed oil, peanut oil, coconut oil, corn oil, castor oil, sesame oil, crude palm oil, fancy mutton tallow, sperm oil, olive oil, tristearin, tripalmitin, triolein and trilaurin. Among the fatty acids utilizable for the purpose of this invention are stearic acid, palmitic acid, oleic acid, linoleic acid and myristic acid.

The source of nitrogenous factors utilizable for the purpose of this invention may be organic (e.g., soybean meal, cornsteep liquor, yeast extract, meat extract and/or distillers' solubles) or synthetic (i.e., composes of simple, synthesizable organic or inorganic compounds, such as ammonium salts, alkali nitrates, amino acids or urea). I

An adequate sterile air supply should be maintained during fennentation, for example, by the conventional limiting) range of the concentration of the compound in the culture is about 0.01 to about 0.1 percent. The culture period (or rather the time of subjecting the compound to the action of the enzyme) may vary considerably, the range of about 24 to 96 hours being feasible, but not limiting.

Compounds A are then acylated to form new Compounds B as by treatment with an excess of an acid anhydride, for example, acetic anhydride in the presence of perchloric acid to yield the l2B,l7a-diacyloxy derivatives. Alternatively, Compounds A may be oxidized directly by treatment with an equivalent amount of an oxidizing agent, such as chromic acid to form the 12-oxo derivatives of Compounds C. This latter compound may then be acylated as set forth above to form the 12-oxo, 17a-acyloxy derivative of Compounds C.

The invention may be illustrated by the following examples, all temperatures are in degrees centigrade unless otherwise stated:

7 EXAMPLE 1 1 23,1 7a-Dihydroxy-A-norprogesterone A. Fermentation Surface growth from each of 3- two-week old agar slants of Corticium microsclerotia (NRRL-2727 the slants containing as a nutrient medium A Oatmeal 20 grams Tomato paste 20 grams Agar 15 grams Tap water to 1 liter is suspended in 6 ml. of 0.01 percent aqueous sodium lauryl sulfate solution. Three ml. portions of this suspension are used to inoculate six 250 ml. Erlenmeyer flasks, each containing 50 ml. of the following sterilized medium (B):

Glucose 30 grams Soy bean meal 20 grams Soy bean oil 2.2 grams CaCO 2.5 grams Distilled water to 1 liter After seventy-two hours incubation at 25 C. with continuous rotary agitation (280 cycles/minute; two inch radium), 10 percent (vol./vol.) transfers are made to thirty-four 250 ml. Erlenmeyer flasks each containing 50 ml. of the following sterilized medium (C):

Dextrose 10 grams Corn steep liquor 6 grams NHJ-I PO 3 grams Yeast extract 2.5 grams CaCO 2.5 grams Distilled water to 1 liter '4 c.p.s., l2a-H) and 4.25 (s, 3-H).

ANAL. Calcd for c u o 332.42 c, 72.26; H,

8.49 Found: c, 72.27; H, 8.33.

EXAMPLE 2 1 2B, 17a-Diacetoxy-A-norprogesterone A solution of 0.0033 ml. of perchloric acid in 0.3 ml. of acetic anhydride is added to 500 mg. of 12,13,170:- dihydroxy-A-norprogesterone in 10 ml. of acetic anhydride. The reaction mixture is stirred at room temperature for 0.5 hours, and then poured into ice-water and extracted with chloroform. The chloroform extracts are washed with a saturated sodium bicarbonate solution, 8 percent salt solution, dried over. sodium sulfate and evaporated to dryness to give 1213,1701- diacetoxy-A-norprogesterone.

EXAMPLE 3 l 2-Ketol 7a-hydroxy-A-norprogesterone A solution of 30 mg. of 12B,17a-dihydroxy-A-norprogesterone in 4 ml. of acetone is treated dropwise with an equivalent amount of chromium trioxide-sulfuric acid. The chromic sulfate is removed by filtration, and washed with additional acetone. The filtrate is concentrated, diluted with water and extracted with chloroform. The chloroform extracts are washed with 8 percent salt solution, dried over sodium sulfate and evaporated to dryness. Crystallization of the residue from chloroforrn-isopropyl ether gives 15 mg. of 12- ketol 7a-hydroxy-A-norprogesterone having a melting point of 178179. The analytical sample is prepared by recrystallization from chloroform-isopropyl ether, m.p. 180.518l.5, M 2.92, 5.88, and 6.17;; A 231 mp.( 15,700).

ANAL. Calcd for C l-1 0 (330.41): C, 72.70; H,

7.93 Found: C, 72.80; H, 8.16.

EXAMPLE 4 1 Z-Keto- 1 7a-acetoxy-A-norprogesterone Following the procedure in Example 2 but substituting 7 r y-Aoqrntq ssterqne o 1213, 17a-dihydroxy-A-norprogesterone there is obtained l2-ket0- 17-acetoxy-A-norprogesterone.

While there have been described various embodiments of the invention, the compositions and methods described are not intended to be understood as limiting the scope of the invention, as it is realized that changes therein are possible and it is further intended that each element recited in any of the following claims is to be understood as referring to all equivalent elements for accomplishing substantially the same results in substan- 5 6 tially the same or equivalent manner, it covering the inwherein R and R are selected from the group consistvention broadly in whatever form its principle may be ing of hydrogen and acyl wherein the acyl is from a tili d, 7 hydrocarbon carboxylic acid of less than twelve carbon What is claimed is: atoms. 1. A compound selected from the group consisting of 5 2. 123,17a-Dihydroxy-A-norprogesterone. the formulas 3. l25,17a-Diacetoxy-A-norprogesterone.

4. lZ-Keto-l7a-hydroxy-A-norprogesterone. on] (1H3 5 l2-Ket0-l7a-acet0x -A-nor r esterone R =0 0 (:0 y P g I... ll l l0 and 

2. 12 Beta ,17 Alpha -Dihydroxy-A-norprogesterone.
 3. 12 Beta ,17 Alpha -Diacetoxy-A-norprogesterone.
 4. 12-Keto-17 Alpha -hydroxy-A-norprogesterone.
 5. 12-Keto-17 Alpha -acetoxy-A-norprogesterone. 